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Ke E , Patel BB , Liu T , Li XM , Haluszka O , Hoffman JP , Ehya H , Young NA , Watson JC , Weinberg DS , Nguyen MT , Cohen SJ , Meropol NJ , Litwin S , Tokar JL , Yeung AT
Proteomic analyses of pancreatic cyst fluids
Pancreas. 2009 Mar;38(2) :e33-42
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OBJECTIVES: There are currently no diagnostic indicators that are consistently reliable, obtainable, and conclusive for diagnosing and risk-stratifying pancreatic cysts. Proteomic analyses were performed to explore pancreatic cyst fluids to yield effective diagnostic biomarkers. METHODS: We have prospectively recruited 20 research participants and prepared their pancreatic cyst fluids specifically for proteomic analyses. Proteomic approaches applied were as follows: (1) matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry peptidomics with LC/MS/MS (HPLC-tandem mass spectrometry) protein identification; (2) 2-dimensional gel electrophoresis; (3) GeLC/MS/MS (tryptic digestion of proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by LC/MS/MS). RESULTS: Sequencing of more than 350 free peptides showed that exopeptidase activities rendered peptidomics of cyst fluids unreliable; protein nicking by proteases in the cyst fluids produced hundreds of protein spots from the major proteins, making 2-dimensional gel proteomics unmanageable; GeLC/MS/MS revealed a panel of potential biomarker proteins that correlated with carcinoembryonic antigen (CEA). CONCLUSIONS: Two homologs of amylase, solubilized molecules of 4 mucins, 4 solubilized CEA-related cell adhesion molecules (CEACAMs), and 4 S100 homologs may be candidate biomarkers to facilitate future pancreatic cyst diagnosis and risk-stratification. This approach required less than 40 microL of cyst fluid per sample, offering the possibility to analyze cysts smaller than 1 cm in diameter.
Ke, Eileen Patel, Bhavinkumar B Liu, Tiffany Li, Xin-Ming Haluszka, Oleh Hoffman, John P Ehya, Hormoz Young, Nancy A Watson, James C Weinberg, David S Nguyen, Minhhuyen T Cohen, Steven J Meropol, Neal J Litwin, Samuel Tokar, Jeffrey L Yeung, Anthony T CA119242/CA/NCI NIH HHS/United States P30CA06927/CA/NCI NIH HHS/United States Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't United States Pancreas Pancreas. 2009 Mar;38(2):e33-42.